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Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy
Published on: December 17, 2019
Co-opting signalling molecules enables logic-gated control of CAR T cells
Aidan M Tousley1, Maria Caterina Rotiroti1, Louai Labanieh2,3
1Department of Pediatrics, Stanford University School of Medicine, Stanford, CA, USA.
Abstract:
Although chimeric antigen receptor (CAR) T cells have altered the treatment landscape for B cell malignancies, the risk of on-target, off-tumour toxicity has hampered their development for solid tumours because most target antigens are shared with normal cells1,2. Researchers have attempted to apply Boolean-logic gating to CAR T cells to prevent toxicity3-5; however, a truly safe and effective logic-gated CAR has remained elusive6. Here we describe an approach to CAR engineering in which we replace traditional CD3ζ domains with intracellular proximal T cell signalling molecules. We show that certain proximal signalling CARs, such as a ZAP-70 CAR, can activate T cells and eradicate tumours in vivo while bypassing upstream signalling proteins, including CD3ζ. The primary role of ZAP-70 is to phosphorylate LAT and SLP-76, which form a scaffold for signal propagation. We exploited the cooperative role of LAT and SLP-76 to engineer logic-gated intracellular network (LINK) CAR, a rapid and reversible Boolean-logic AND-gated CAR T cell platform that outperforms other systems in both efficacy and prevention of on-target, off-tumour toxicity. LINK CAR will expand the range of molecules that can be targeted with CAR T cells, and will enable these powerful therapeutic agents to be used for solid tumours and diverse diseases such as autoimmunity7 and fibrosis8. In addition, this work shows that the internal signalling machinery of cells can be repurposed into surface receptors, which could open new avenues for cellular engineering.
Insights
Researchers developed a novel logic-gated intracellular network (LINK) CAR T cell platform. This innovative approach enhances anti-tumour efficacy while preventing on-target, off-tumour toxicity, paving the way for solid tumour treatments.
Area of Science:
- Immunology
- Cellular Engineering
- Cancer Therapy
Background:
- Chimeric antigen receptor (CAR) T cells are effective against B cell malignancies but face toxicity challenges in solid tumours due to shared antigens.
- Existing logic-gated CAR T cell strategies have not fully resolved safety and efficacy issues.
- Targeting solid tumours with CAR T cells requires overcoming on-target, off-tumour toxicity.
Purpose of the Study:
- To engineer a novel CAR T cell platform with enhanced safety and efficacy for solid tumour treatment.
- To develop a reversible Boolean-logic AND-gated CAR T cell system.
- To overcome limitations of current CAR T cell therapies by bypassing upstream signalling molecules.
Main Methods:
- Replaced traditional CD3ζ domains in CARs with intracellular proximal T cell signalling molecules.
- Engineered a logic-gated intracellular network (LINK) CAR T cell platform utilizing ZAP-70, LAT, and SLP-76 signalling.
- Evaluated the efficacy and toxicity profile of the LINK CAR T cells in preclinical models.
Main Results:
- Demonstrated that proximal signalling CARs, like ZAP-70 CAR, can activate T cells and eradicate tumours in vivo.
- The LINK CAR platform functions as a rapid and reversible Boolean-logic AND-gated system.
- LINK CAR T cells exhibited superior efficacy and prevention of on-target, off-tumour toxicity compared to other systems.
Conclusions:
- The LINK CAR platform represents a significant advancement in CAR T cell engineering, offering improved safety and efficacy.
- This technology expands the potential targets for CAR T cell therapy, including solid tumours and autoimmune diseases.
- Repurposing intracellular signalling machinery into surface receptors opens new possibilities for cellular engineering.
